US4536407A - Functional protein products - Google Patents
Functional protein products Download PDFInfo
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- US4536407A US4536407A US06/518,593 US51859383A US4536407A US 4536407 A US4536407 A US 4536407A US 51859383 A US51859383 A US 51859383A US 4536407 A US4536407 A US 4536407A
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- protein product
- cells
- functional protein
- emulsion
- foodstuff
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 56
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 56
- 210000004027 cell Anatomy 0.000 claims abstract description 65
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 34
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 34
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 34
- 210000005253 yeast cell Anatomy 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims description 31
- 239000006071 cream Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 239000000839 emulsion Substances 0.000 claims description 23
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 20
- 239000002585 base Substances 0.000 claims description 20
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 19
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 17
- 230000000813 microbial effect Effects 0.000 claims description 11
- 241000235648 Pichia Species 0.000 claims description 9
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 8
- 239000003086 colorant Substances 0.000 claims description 8
- 241000235058 Komagataella pastoris Species 0.000 claims description 7
- 239000002253 acid Substances 0.000 claims description 6
- 241000235646 Cyberlindnera jadinii Species 0.000 claims description 5
- 241000894007 species Species 0.000 claims description 5
- 241000235070 Saccharomyces Species 0.000 claims description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 claims description 3
- 241001149669 Hanseniaspora Species 0.000 claims description 3
- 241000235649 Kluyveromyces Species 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 241000222178 Candida tropicalis Species 0.000 claims description 2
- 235000014663 Kluyveromyces fragilis Nutrition 0.000 claims description 2
- 241000320412 Ogataea angusta Species 0.000 claims description 2
- 241000235059 Ogataea pini Species 0.000 claims description 2
- 241000222124 [Candida] boidinii Species 0.000 claims description 2
- 239000008157 edible vegetable oil Substances 0.000 claims 7
- 239000007900 aqueous suspension Substances 0.000 claims 4
- 238000005406 washing Methods 0.000 claims 4
- 150000008044 alkali metal hydroxides Chemical class 0.000 claims 2
- 241000235650 Kluyveromyces marxianus Species 0.000 claims 1
- 230000002378 acidificating effect Effects 0.000 abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 27
- 239000000047 product Substances 0.000 description 17
- 239000006260 foam Substances 0.000 description 14
- 239000007788 liquid Substances 0.000 description 13
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 12
- 230000004151 fermentation Effects 0.000 description 12
- 239000000463 material Substances 0.000 description 12
- 238000000855 fermentation Methods 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 230000001413 cellular effect Effects 0.000 description 8
- 230000001804 emulsifying effect Effects 0.000 description 8
- 239000000758 substrate Substances 0.000 description 8
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000006285 cell suspension Substances 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000000725 suspension Substances 0.000 description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 4
- 239000000908 ammonium hydroxide Substances 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000006185 dispersion Substances 0.000 description 4
- 229910052500 inorganic mineral Inorganic materials 0.000 description 4
- 235000010755 mineral Nutrition 0.000 description 4
- 239000011707 mineral Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 108010027322 single cell proteins Proteins 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 235000013619 trace mineral Nutrition 0.000 description 4
- 239000011573 trace mineral Substances 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 108010073771 Soybean Proteins Proteins 0.000 description 3
- 238000010306 acid treatment Methods 0.000 description 3
- -1 aliphatic hydrocarbyl alcohols Chemical class 0.000 description 3
- 229910021529 ammonia Inorganic materials 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 125000004432 carbon atom Chemical group C* 0.000 description 3
- 230000009089 cytolysis Effects 0.000 description 3
- 238000005187 foaming Methods 0.000 description 3
- 239000013505 freshwater Substances 0.000 description 3
- 229930195733 hydrocarbon Natural products 0.000 description 3
- 150000002430 hydrocarbons Chemical class 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000005086 pumping Methods 0.000 description 3
- 230000035939 shock Effects 0.000 description 3
- 229940001941 soy protein Drugs 0.000 description 3
- 238000001694 spray drying Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000012736 aqueous medium Substances 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 239000002285 corn oil Substances 0.000 description 2
- 235000005687 corn oil Nutrition 0.000 description 2
- 238000002635 electroconvulsive therapy Methods 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 235000015074 other food component Nutrition 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000010935 stainless steel Substances 0.000 description 2
- 241000191291 Abies alba Species 0.000 description 1
- 102100034612 Annexin A4 Human genes 0.000 description 1
- 108090000669 Annexin A4 Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 101710161822 Extracellular ribonuclease Proteins 0.000 description 1
- 229910003556 H2 SO4 Inorganic materials 0.000 description 1
- 244000285963 Kluyveromyces fragilis Species 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000010564 aerobic fermentation Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 229910000272 alkali metal oxide Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical class 0.000 description 1
- 229910000287 alkaline earth metal oxide Inorganic materials 0.000 description 1
- 238000010945 base-catalyzed hydrolysis reactiony Methods 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 150000004649 carbonic acid derivatives Chemical class 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000000306 component Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007797 corrosion Effects 0.000 description 1
- 238000005260 corrosion Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 229910001882 dioxygen Inorganic materials 0.000 description 1
- 238000002036 drum drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 235000021474 generally recognized As safe (food) Nutrition 0.000 description 1
- 235000021473 generally recognized as safe (food ingredients) Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 125000000864 peroxy group Chemical class O(O*)* 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000012047 saturated solution Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000002195 soluble material Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/18—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from yeasts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS
- A23D9/00—Other edible oils or fats, e.g. shortenings or cooking oils
- A23D9/007—Other edible oils or fats, e.g. shortenings or cooking oils characterised by ingredients other than fatty acid triglycerides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/82—Proteins from microorganisms
- Y10S530/821—Separation of nucleic acid
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/82—Proteins from microorganisms
- Y10S530/823—Lower fungi, e.g. mold
- Y10S530/824—Yeasts
Definitions
- the invention pertains to whippable protein products.
- the invention pertains to methods to produce a whippable protein product from microbial cells.
- the invention pertains to whipped nucleic acid-reduced proteins.
- a protein product for commercial acceptance should have good functional properties as well as be suitable for human consumption.
- Microbial cells have high potential as protein sources, being relatively cheaply grown on a wide variety of substrates.
- nucleic acid content of microbial protein initially is high.
- Such proteins unless reduced in nucleic acid content can be used by humans only in very limited amounts.
- a variety of means have been developed for reduction in nucleic acid content.
- nucleic acid-reduced protein products still need acceptable functional properties, such as whippability, before broad acceptance in the market place as a food qualtity product.
- step (C) whipping the treated whole cells from step (B); or less preferably the soluble fraction from step (B).
- the product resulting from this series of steps is comestible, has very low nucleic acid content, and exhibits good functionality, i.e., whippability.
- Functional properties is a collective term for those physicochemical properties of proteins which govern their composite performance in foods during manufacturing, processing, storage, and consumption, reflecting the properties of the protein that are influenced by its composition, conformation, and interactions with other food components.
- Functional properites include the important properties of whippability and foam capability and stability, Kinsella, J. E. "Functional Properties of Proteins in Foods: A Survey”, CRC Critical Reviews in Food Sci. and Nutr. 7, 219 (1976).
- yeasts are employed.
- yeasts can be grown on a suitable carbon energy source (substrate), under aerobic aqueous fermentation conditions employing assimilable nitrogen-source, mineral salts aqueous media, molecular oxygen, with suitable pH and other controls, all as known in the art.
- the carbon energy substrate can be any carbon energy source, such as hydrocarbons, oxygenated hydrocarbons, including various carbohydrates, and the like, suitable as yeast substrates. It is recognized that particular yeasts do vary in their preference for various substrates.
- the presently preferred substrates for aqueous fermentation conditions are the oxygenated hydrocarbons or carbon-oxygen-hydrogen (C--O--H) compounds which are significantly water-soluble.
- C--O--H is intended to be a generic term in this disclosure descriptive of compounds employable, and not necessarily a limiting term referring to the source of the substrate.
- the C--O--H compounds include the water-soluble carbohydrates, as well as those alcohols, ketones, esters, acids, and aldehydes, and mixtures, which are reasonably significantly water-soluble in character, generally of 1 to 20 carbon atoms per molecule.
- Preferred are the C--O--H compounds which exhibit the greater water-solubility.
- Preferred are the water-soluble linear monohydric aliphatic hydrocarbyl alcohols, particularly methanol and ehtanol.
- yeast strains are chosen from the following genera: Saccharomyces, Candida, Torulopsis, Pichia, Hansenula, Kluyveromyces, and Kloeckera.
- Exemplary species include:
- yeasts of the Candida, Saccharomyces, and Pichia genera of these particularly, Saccharomyces cerevisiae, Candida utilis, and Pichia pastoris, particularly such as it Pichia pastoris NRRL Y-11430 and Y-11431.
- Yeast cells in general can be employed, including pre-produced, separated from aqueous ferment, and/or dried. Fresh cells from a continuous producing fermentation preferably are employed.
- Nucleic acid reduction should be accomplished by a method that avoids significant lysis of the cells.
- the aqueous ferment (suspended cells plus aqueous medium) exiting the fermentation step contains both supernatant aqueous liquor and suspended cells.
- the cells can be separated, by such as centrifugation or filtering, water-washed if desired, and resuspended in water.
- the cell concentration in the aqueous ferment can be increased, if needed, by vacuum filtration, solvent removal, or the like. If the aqueous ferment is from a high cellular density fermentation, it can be used as is in some nucleic acid removal processes.
- One suitable process treats the cells without significant lysis as a cell cream containing a solids content of about 10 to 25, preferably 12 to 20, weight percent (dry basis) with a base such as NH 4 OH to a pH of such as about 7 to 10, preferably 8.5 to 10, accompanied with heating to a temperature in the range of about 65° C. to 99° C., preferably 85° C. to 99° C., for a base-treating time of about 15 to 120 minutes, preferably 30 to 60 minutes. Thereafter, the cells are separated, and water-washed. While the cells remain substantially intact in this step, the base treatment step reduces substantially the nucleic acid content of the cells.
- Suitable bases include ammonia, and ammonium and water soluble alkali metal and alkaline earth metal oxides, hydroxides, and carbonates, such as ammonium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, potassium carbonate, and the like, and mixtures thereof.
- the bases normally are employed in the form of water solutions, in as concentrated a form as is convenient. Thus, it is preferred to employ ammonia, or NH 4 OH as a commercially available concentrated solution of about 28 weight percent.
- nucleic acid reduction Any other methods of nucleic acid reduction can be used, so long as the method employed is effective in reducing the nucleic acid content of the cells, without appreciable rupture of the cells (such that appreciable lysis is not effected). It is realized that in any of the methods employed that some cellular damage may occur to some of the cells, with some protein entering the aqueous treating solution. Such solubilized or extruded proteins can be separated and used for other food purposes.
- Typical suitable methods include: treatment of cells with ammonia or ammonium hydroxide with an alcohol such as methanol, South Africa 774517; treatment of cells with extra-cellular ribonuclease, U.S. Pat. No. 3,960,659; degradation of nucleic acids with endogenous nucleases, U.S. Pat. No. 3,809,776; treatment of cells with anhydrous liquid ammonia, U.S. Pat. No. 3,615,654; other methods for removal of nucleic acids from single cell protein materials as described in Single Cell Protein II, S. R. Tannenbaum and D. I. C.
- nucleic acid removal is accomplished by a first acid- or base-treatment step
- a second treatment of opposite pH i.e., acidic wash followed by base wash or base wash followed by acidic wash
- acid treatment is to be employed following an initial base treatment
- a pH of about 3 to 5 for 20 to 50 minutes at 80° C.-95° C. typically is employed.
- the treated cells preferably are separated, such as by centrifugation, from the aqueous treating liquor.
- separated nucleic acid reduced cells are washed with such as about 1 to 4 volumes of water per volume of packed cells to remove essentially all traces of treating chemicals prior to the next step in the process of my invention, to aid removal of solubilized but adsorbed or adsorbed nucleic acids and to avoid potential equipment corrosion.
- the nucleic acid-reduced cells are re-admixed with sufficient water to form a cell cream of a cellular concentration (solids content) of about 10 to 25, preferably 12 to 20, weight percent (dry basis).
- the cellular cream then is subjected to a short-time heat-shock at a temperature of about 110° C. to 160° C., preferably about 120° C. to 140° C., for a time of about 40 to 200 seconds, preferably about 80 to 120 seconds.
- the heat-shock step can be accomplished by any suitable means such as by pumping the cream through a heated coil or tubing or heat exchanger sufficient to attain the elevated temperature short-time heat-shock treatment. Pressure is not considered critical, and the cream can be pumped therethrough at pressures as convenient, such as about 10 to 100 psig, preferably 20 to 50 psig.
- a 1/4" to 3/8" ID stainless steel tubing coil externally heated by high pressure steam such as at 30 psig and 140° C., including convenient means to vary the tubing length and pumping rate to achieve the desired residence time can be used to provide a suitable throughput.
- the residence time required to achieve a product with improved functional properties typically will be less for a higher temperature treatment than the residence time required for a lower temperature treatment, within the temperature range indicated above.
- a cream residence time in the heated zone of about 60 to 120 seconds.
- nucleic acid removal and heat shock to impart improved functional properties (whippability) to the protein product can be carried out coincidentally, i.e., essentially at the same time.
- a crude fermentation effluent can be adjusted to pH of about 9 to 11 with a suitable base as described previously, and then subjected to heat shock conditions as described above.
- the so-treated effluent can then be separated into (a) an insoluble cellular fraction of reduced nucleic acid content, and (b) a liquid fraction which also will have good functional properties though containing much of the nucleic acids.
- the heat-shocked cells from step (B), or the cellular fraction from step (A-B), preferably are reduced in water content, and, if desired, dried.
- Reduction in water content and/or drying can be accomplished by spray drying, drum drying, freeze drying, or the like.
- spray drying can be employed using an inlet temperature of such as about 540° F., and an outlet temperature of such as about 210° F.
- the product of my process is a comestible, functional, protein product exhibiting good whippability, forming a highly stable foam. This property is important in use of the protein in food products to provide good mixing, particularly in preparing light, fluffy comestibles with minimum weights of ingredients.
- the product is mixed with water, and whipped or beaten, such as in a blender, a sufficient speed and under conditions effective to admix air therewith to produce a foam of desired thickness, texture, creaminess, and gaseous content.
- gases can be used, if desired, such as nitrogen.
- Colorants, or GRAS character can be employed to impart tints or colorations as may be desired.
- Two or more foams of differing tints can be lightly mixed to produce stable multi-colored (parti-colored) foam swirls.
- the whipped material can be incorporated into various food-stuffs after whipping. Or, the basic product can be incorporated with other food components and then whipped. In either case, the whipped material adds volume, stability, good quality protein, at low weight and low-cost.
- methanol and an aqueous mineral salts medium in a volume ratio of about 40 to 60, respectively, were fed individually to a fermentor, inoculated with the yeast species Pichia pastoris NRRL Y-11430, at a rate such that methanol was the growth-limiting factor.
- the fermentor was a 1500-liter foam-filled fermentor with a liquid volume of about 610 liters, with automatic pH, temperature, and level control. Agitation was provided by two conventional paddle-type turbines driven at 1000 rpm. The aeration rate was about 4 volumes of air (at about 38 psig and about 25° C.) per volume of ferment in the fermentor per minute. Anhydrous ammonia was added at a rate sufficient to maintain a pH of about 3.5 in the fermentation mixture.
- the aqueous mineral salts medium was prepared by mixing, with each liter of tap water, 15.86 mL 75 wt percent H 3 PO 4 ,9.53 g K 2 SO 4 , 7.8 g MgSO 4 .7H 2 O, 0.6 g CaCl 2 . 2H 2 O, and 2.6 g 85 wt percent KOH.
- the aqueous mineral salts medium was fed at a rate of 31.5 liters per hour and the methanol at a rate of 21 liters per hour.
- the trace mineral solution was prepared by mixing, for each liter of solution 65 g FeSO 4 .7H 2 O, 20 g ZnSO 4 .7H 2 O, 3.0 g MnSO 4 .H 2 O, 6.0 g CuSO 4 .5H 2 O, 5.0 mL conc. H 2 SO 4 , and sufficient deionized water to make 1 liter of solution.
- the trace mineral solution plus biotin was prepared by mixing 780 mL of a trace mineral solution, 20 mL water, 200 mL methanol and 0.032 g biotin. The trace mineral solution plus biotin was fed separately via the methanol stream at a rate of 10 mL per liter of methanol.
- the fermentation was conducted at about 30° C. and about 38 psig pressure, with an average retention time of about 11.6 hours.
- the cell density typically was about 128.4 g of cells per liter of fermentor effluent.
- the total solids content of the ferment typically was about 134.7 g per liter.
- the resulting yeast cells were separated from the fermentation effluent (aqueous ferment) by centrifugation, washed by suspension in water, followed by recentrifugation, dried overnight at 100° C., and weighed. On a dried basis, the yield of yeast cells typically was about 40.6 g per 100 g of methanol fed.
- NARP Nucleic Acid Reduced SCP
- Fermentor effluent as obtained from runs as described in Example I, was adjusted to a pH of 9.0 to 9.5 with concentrated NH 4 OH solution, then heated to 90° C. to 95° C. for 20 to 40 minutes.
- the resulting cellular dispersion (suspension) was centrifuged, and the liquid removed and discarded.
- the cell paste was resuspended in about 4 volumes fresh water, then centrifuged again. The supernatant liquid was discarded.
- the cell paste was collected for further processing. This material was designated as nucleic acid reduced protein (NARP).
- NARP nucleic acid reduced protein
- the NH 4 OH treated cells prepared as described in Example II, were resuspended in water to a concentration of about 12 to 20 wt percent, and the dispersion adjusted to a pH of 4.2 with sulfuric acid. The dispersion was heated to 85° C. for 30 minutes. The resulting dispersion was centrifuged, and the liquid removed and discarded. The cell paste was resuspended in about 4 volumes fresh water, again centrifuged, and the supernatant liquid discarded. The cell paste was collected for further processing. This acid-treated material was designated as NARP (AS).
- AS NARP
- NARP prepared as described in Example II
- the washed cells were resuspended in water to a concentration of about 12 to 20 wt percent, then treated with 2 mL of 30% H 2 O 2 per liter of total cell suspension and heated to 60° C. for 15 minutes.
- the resulting suspension was centrifuged, and the liquid removed and discarded.
- the cell paste was resuspended in about 4 volumes fresh water, again centrifuged, and the supernatant liquid discarded.
- the cell paste was collected for further processing.
- This ammonium hydroxide and peroxy-treated material was designated as NARP (AO).
- the heat-shock step typically was carried out employing a cell cream containing from 120 to 200 g of cells/L (dry basis). More concentrated cream tends to plug the tubing at the operating conditions employed, while cream less concentrated than 120 g/L is less efficient than the typical operating range employed.
- the cream employed was the washed effluent from a prior acid- or base-treatment.
- a heat-shocked cream typically had a pH in the range of about 5-8.5.
- the cell suspension (cream) was pumped via stainless steel tubing (1/4" 316SS) through a coil of tubing (10 foot length, 3" internal coil) placed in a silicon oil bath.
- the oil bath was maintained at about 140° C., reaction pressure of about 30 psig employed, and residence time of cell suspension in the heat bath adjusted to about 80 seconds by varying the pumping rate and/or the length of the first tubing coil.
- the tubing carried the cellular cream suspension through a second coil of tubing placed in an ice-bath to accomplish rapid cooling of the cell suspension.
- the product was dried for feeding studies by lyophilizing or by employing a spray dryer.
- Cells to be lyophilized were cooled at about -100° F. to -150° F. and pressure reduced to about 20 milli Torr, for about 6 to 24 hours as necessary to accomplish complete sample drying.
- Spray drying was accomplished by feeding the treated cream into a rotating drum with filtered inlet air at about 540° F. Liquid feed rate was maintained so as to achieve an outlet air temperature of about 210° F. Under typical conditions, about 18 gallons/hour of cream were pumped through the spray dryer.
- the whippability character of each of several different protein materials was measured by suspending 5 g of protein in 50 mL of water in a stoppered 100 mL graduated cylinder. Samples were shaken vigorously for one minute, then shaking discontinued, and the liquid and foam levels were measured at intermittent time intervals. Table I presents foam volumes over liquid volumes in mL:
- Emulsifying ability was measured. Protein (1.75 g) was placed in a 50 mL graduated cylinder, and water added to give a total volume of 20 mL, mixed well to give a suspension, and then poured into a Waring Blender jar. The gradulated cylinder was rinsed with 5 mL of water, which also was added to the blender jar to ensure complete transfer of the protein material. Mazola corn oil (25 mL) then was added to the blender jar, and the mixture blended at maximum speed for 1 minute. A portion of the resultant emulsion was poured into a centrifuge tube and spun for 5 minutes at 2000 rpm. The volume of the emulsified layer was divided by the total volume of mixture multiplied by 100 to determine the Emulsifying Index of the sample. Results are shown in Table II:
- NARP(AS) (HS) (from Run 4) has emulsion-forming capabilities comparable to soy protein.
- the emulsion from the protein of Run 4 was a whiter, denser, and thicker emulsion.
- the stability of the emulsions obtained above was tested by adjusting the emulsion pH to basic or acidic pH until the emulsion broke.
- the NARP(AS) (HS) emulsion of Run 4 had an initial pH of ⁇ 4.3 and withstood a pH of up to 12.5 before the emulsion began to liquefy, a greater stability than the others tested.
- Fermentor effluent prepared as described in Example I, was adjusted to a pH of about 9.5 with a saturated solution of NaOH, then subjected to heat-shock conditions (130° C. for 2 minutes) employing the apparatus described above in Example V.
- the treated cell suspension was separated by centrifugation into a solid and liquid fraction.
- a sample of the solid phase was analyzed for protein and nucleic acid content and found to have 58 wt percent protein and a nucleic acid content of 2.0 wt percent.
- a sample of the liquid phase was analyzed for protein and nucleic acid content.
- the sample was lyophilized giving about 55 g/L of solubles.
- the protein content of the dried solubles was about 49 wt percent and nucleic acid content about 3.9 wt percent.
- Emulsifying ability was determined as described in Example VII, except that 3.5 g of lyophilized heat-shocked soluble material, 50 mL of water, and 50 mL of Mazola corn oil were employed. The average emulsifying index obtained from 6 separate determinations was 85%.
- the "solubles" product exhibits good emulsifying characteristics. However, the foam quality and stability were below that of the foam of Run 4 (See Table I). Further, the nucleic acid content is higher than desirable, though tolerable in usages where smaller quantities are acceptable.
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- Food Science & Technology (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
______________________________________
Saccharomyces cerevisiae
Torulopsis methanosorbosa
Candida boidinii Torulopsis methanothermo
Candida methanolica
Pichia lindnerii
Candida tropicalis Pichia pastoris
Candida utilis Pichia pinus
Torulopsis methanolovescens
Hansenula polymorpha
Torulopsis methanophiles
Kluyveromyces fragilis
______________________________________
TABLE I
______________________________________
Time, min.
Run # 1 2 5 15 30
______________________________________
1 Whole Cell 60 55 51 47 46
26 30 34 37 38
2 NARP 21 20 18 17 16
40 41 42 43 43
3 NARP(AS) 34 32 29 26 26
38 40 41 44 44
4 NARP(AS)(HS)
83 76 71 67 65
21 28 32 35 37
5 NARP(AO)(HS)
32 31 30 29 28
37 38 39 40 41
6 NBC Soy Protein
34 33 31 27 23
41 42 44 46 48
______________________________________
TABLE II
______________________________________
Total Emulsion
Volume, Volume, Emulsifying
Sample mL mL Index, %
______________________________________
Whole Cell (Run 1)
14.7 9.4 63.9
NARP(AS)(HS) (Run 4)
14.5 8.1 55.9
NBC Soy Protein (Run 6)
14.8 8.0 54.1
______________________________________
TABLE III ______________________________________ Time, min. 1 2 5 15 30 ______________________________________ 92 85 73 57 31 13 20 32 43 49 ______________________________________
Claims (31)
______________________________________
Saccharomyces cerevisiae,
Torulopsis methanosorbosa,
Candida boidinii, Torulopsis methanothermo,
Candida methanolica,
Pichia lindnerii,
Candida tropicalis,
Pichia pastoris,
Candida utilis, Pichia pinus,
Torulopsis methanolovescens,
Hansenula polymorpha, and
Torulopsis methanophiles,
Kluyveromyces fragilis.
______________________________________
Priority Applications (2)
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| US06/518,593 US4536407A (en) | 1983-07-29 | 1983-07-29 | Functional protein products |
| US06/713,506 US4564523A (en) | 1983-07-29 | 1985-03-18 | Functional protein products |
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| US06/518,593 US4536407A (en) | 1983-07-29 | 1983-07-29 | Functional protein products |
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| US06/713,506 Division US4564523A (en) | 1983-07-29 | 1985-03-18 | Functional protein products |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4904485A (en) * | 1986-10-02 | 1990-02-27 | Kanegafuchi Kagaku Kogyo Kabushiki Kaisha | Fat compositions suitable for use in bakeries or confectioneries |
| US5381914A (en) * | 1991-05-09 | 1995-01-17 | Toyo Seikan Kaisha, Ltd. | Container closure with liner |
| US5914345A (en) * | 1994-10-11 | 1999-06-22 | Endoluminal Therapeutics, Inc. | Treatment of tissues to reduce subsequent response to injury |
| CN118020921A (en) * | 2021-11-16 | 2024-05-14 | 上海昌进生物科技有限公司 | Composition, preparation method and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4904485A (en) * | 1986-10-02 | 1990-02-27 | Kanegafuchi Kagaku Kogyo Kabushiki Kaisha | Fat compositions suitable for use in bakeries or confectioneries |
| US5381914A (en) * | 1991-05-09 | 1995-01-17 | Toyo Seikan Kaisha, Ltd. | Container closure with liner |
| US5914345A (en) * | 1994-10-11 | 1999-06-22 | Endoluminal Therapeutics, Inc. | Treatment of tissues to reduce subsequent response to injury |
| US6071956A (en) * | 1994-10-11 | 2000-06-06 | Endoluminal Therapeutics, Inc. | Treatment of tissues to reduce subsequent response to injury |
| CN118020921A (en) * | 2021-11-16 | 2024-05-14 | 上海昌进生物科技有限公司 | Composition, preparation method and application thereof |
| JP2024542215A (en) * | 2021-11-16 | 2024-11-13 | 上海昌▲進▼生物科技有限公司 | Compositions, methods for preparing same and uses |
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